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Nutrients 2018, 10(6), 757; https://doi.org/10.3390/nu10060757

Immunological Comparison of Native and Recombinant Hen’s Egg Yolk Allergen, Chicken Serum Albumin (Gal d 5), Produced in Kluveromyces lactis

1
NeuroAllergy Research Laboratory (NARL), School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia
2
Poultry Cooperative Research Centre, P.O. Box U242, University of New England, Armidale, NSW 2351, Australia
3
Department of Biological Science, Faculty of Applied Sciences, Rajarata University of Sri Lanka, Mihintale 50300, Sri Lanka
4
Australian Animal Health Laboratory (AAHL), Biosecurity Flagship, Commonwealth Scientific and Industrial Research Organisation (CSIRO), 5 Portarlington Road, Newcomb, VIC 3219, Australia
5
Department of Allergy and Immunology, Royal Children’s Hospital, 50 Flemington Road, Parkville, VIC 3052, Australia
6
Allergy and Immune Disorders, Murdoch Children’s Research Institute, Parkville, Melbourne, Victoria 3052, Australia
7
Department of Paediatrics, The University of Melbourne, Parkville, Melbourne, Victoria 3052, Australia
*
Author to whom correspondence should be addressed.
Received: 17 May 2018 / Revised: 5 June 2018 / Accepted: 8 June 2018 / Published: 12 June 2018
(This article belongs to the Special Issue Contributions of Diet and Gastrointestinal Digestion to Food Allergy)
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Abstract

Chicken serum albumin (CSA) is a hen’s egg yolk allergen causing IgE-mediated allergy. The objective of this study was to produce a recombinant version of CSA and compare its IgE reactivity to natural CSA (nCSA). CSA was cloned and expressed as a soluble fraction in the yeast Kluyveromyces lactis (K. lactis) protein expression system. The gene encoding CSA was amplified with a C-terminal hemagglutinin epitope tag by polymerase chain reaction (PCR) and cloned into the pKLAC2 expression vector prior to transforming into K. lactis. Recombinant CSA (rCSA) was purified by immunoprecipitation. Twenty-one patients allergic to hen’s egg white were examined for sensitisation against nCSA. 38% of patients were found to be sensitised to CSA based on Western immunoassay. Immunoglobulin E (IgE) binding capacity of rCSA and nCSA was analysed by ELISA using sera from patients sensitised to CSA. Levels of IgE-binding were similar for both the recombinant and the natural CSA, indicating the existence of similar epitopes. rCSA produced in this study is a potential candidate to be used in component-resolved diagnosis (CRD) of egg yolk allergy. The usefulness of rCSA in CRD of egg yolk allergy warrants further characterisation using sera from patients with allergy to hen’s egg yolk in future studies. View Full-Text
Keywords: allergy; egg yolk allergens; recombinant allergens; chicken serum albumin allergy; egg yolk allergens; recombinant allergens; chicken serum albumin
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De Silva, C.; Dhanapala, P.; King, S.; Doran, T.; Tang, M.; Suphioglu, C. Immunological Comparison of Native and Recombinant Hen’s Egg Yolk Allergen, Chicken Serum Albumin (Gal d 5), Produced in Kluveromyces lactis. Nutrients 2018, 10, 757.

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